Isolation, characterization of Vibrio and Pseudomonas spp from infected fresh water ornamental fishes and evaluation of potential agents for its control

The present work aims at comparing the antibacterial effect of probiotics, plant extracts of Vitex negundo and Clitoria ternatae and antibiotics against the disease causing bacteria isolated from infected ornamental fishes. The molecular characterization of isolated pathogens was performed by randomly amplified polymorphic DNA (RAPD) technique. The antibacterial effects of probiotic and plant extracts were analyzed by well diffusion method and antibiotic disc was performed by disc diffusion method. Isolated probiotics such as Bacillus sp. showed maximum antibacterial activity of 18 mm and 16 mm zone against Vibrio  sp (V2) and Pseudomonas sp (Ps1) respectively. Leaf extract of Vitex negundo displayed zone of inhibition of 20 mm on Pseudomonas sp (Ps3) and 16 mm on Vibrio sp (V1). Among the antibiotics,  chloramphenicol and ampicillin showed maximum inhibitory activity against Vibrio sp (V3) (25mm) and Pseudomonas sp (Ps2) (25mm) respectively. Even though, antibiotics showed higher inhibitory activity  than the isolated Bacillus bacteria and plant extract (V. negundo and C. ternatae). This study concluded that the use of probiotics as an alternative strategy to the use of antibiotics because of its effective  antibacterial activity and growing concern in violence of disinfectants and antibacterial agents

Intracellular granzyme A expression of peripheral blood lymphocyte subsets in pulmonary tuberculosis

Cell-mediated immunity is a key weapon of host defence against tuberculosis (TB). Granzyme A (GzmA), a serine protease, present in the granules of cytotoxic cells induces caspase-independent cell death. We estimated the proportion of GzmA producing lymphocyte subsets in peripheral blood from 59 normal healthy volunteers and 48 pulmonary TB (PTB) patients using flow cytometry. When compared with normal healthy subjects, we observed a significantly higher percentage of GzmA-positive CD56+ cells (P = 0.01) in PTB patients. However, when the absolute number was compared between the two groups, a significantly decreased number of GzmA-expressing CD16+ (P = 0.01) and CD56+ (P = 0.0001) cells was observed in patients and this could be explained by the significantly reduced number of total lymphocytes (P = 0.0009) seen in the patients. There was no significant difference in the number of CD4+ and CD8+ GzmA double-positive cells between the two study groups. CD56 is a natural killer cell marker and these cells represent innate immune response to TB. We report an increased percentage of CD56+ cells expressing GzmA in TB patients, which shows the relevance of the GzmA-mediated pathway of apoptosis in immunity against Mycobacterium tuberculosis

Elevated percentage of perforin positive cells in active pulmonary tuberculosis

Background & objectives: Perforin is one of the major effector molecules of cytotoxic cells associated with killing of cells harbouring intracellular bacterial infection. The precise role of perforin positive cells in tuberculosis still remains controversial. The present study was done to determine the number of circulating CD4+ and CD8+ perforin positive cells to assess the level of cytotoxic response against Mycobacterium tuberculosis in patients with pulmonary tuberculosis. Methods: Intracellular perforin and surface CD4 and CD8 staining of peripheral blood lymphocytes was done using specific monoclonal antibodies and enumerated using flowcytometry. Results: A significantly decreased total lymphocytes (P<0.01), CD4 (P<0.001) and CD8 (P<0.01) lymphocyte counts in PTB patients was observed compared to normal healthy individuals (NHS). Intracellular perforin staining showed significantly elevated percentages of total (P<0.05) and CD8 (P<0.01) perforin positive cells in PTB patients compared to NHS. However, the absolute counts of total, CD4 and CD8 cells positive for perforin were similar in patients and NHS. Interpretation & conclusion: Our results suggest that during active stage of pulmonary tuberculosis there was an increased percentage of CD8 cells positive for perforin, irrespective of their absolute counts. Further, CD8+ perforin positive cells ma

Breast cancer management pathways during the COVID-19 pandemic: outcomes from the UK ‘Alert Level 4’ phase of the B-MaP-C study

Abstract: Background: The B-MaP-C study aimed to determine alterations to breast cancer (BC) management during the peak transmission period of the UK COVID-19 pandemic and the potential impact of these treatment decisions. Methods: This was a national cohort study of patients with early BC undergoing multidisciplinary team (MDT)-guided treatment recommendations during the pandemic, designated ‘standard’ or ‘COVID-altered’, in the preoperative, operative and post-operative setting. Findings: Of 3776 patients (from 64 UK units) in the study, 2246 (59%) had ‘COVID-altered’ management. ‘Bridging’ endocrine therapy was used (n = 951) where theatre capacity was reduced. There was increasing access to COVID-19 low-risk theatres during the study period (59%). In line with national guidance, immediate breast reconstruction was avoided (n = 299). Where adjuvant chemotherapy was omitted (n = 81), the median benefit was only 3% (IQR 2–9%) using ‘NHS Predict’. There was the rapid adoption of new evidence-based hypofractionated radiotherapy (n = 781, from 46 units). Only 14 patients (1%) tested positive for SARS-CoV-2 during their treatment journey. Conclusions: The majority of ‘COVID-altered’ management decisions were largely in line with pre-COVID evidence-based guidelines, implying that breast cancer survival outcomes are unlikely to be negatively impacted by the pandemic. However, in this study, the potential impact of delays to BC presentation or diagnosis remains unknown

Neuromyelitis optica-IgG testing in an Indian cohort with neuromyelitis optica and related demyelinating disorders: Our experience

Background: Neuromyelitis optica (NMO) is an immune-mediated inflammatory demyelinating disorder of the central nervous system with a predilection for the optic nerves and the spinal cord. Immunopathological evidence suggests that the target antigen of the disease is aquaporin-4. An IgG antibody against this protein has been explored as a molecular marker for the disease and as a diagnostic tool due to its high sensitivity and specificity in various populations. Objective: To assess the value of NMO-IgG testing in Indian patients with clinical and magnetic resonance imaging features consistent with NMO and longitudinally extensive transverse myelitis (LETM). Materials and Methods: Forty-five patients with clinical and magnetic resonance imaging features consistent with NMO, LETM, and MS were tested for serum NMO-IgG. Of these patients, 22 patients satisfied revised (2006) Wingerchuk criteria for NMO (excluding NMO-IgG status) and 11 patients had LETM. Twelve patients satisfied the revised (2010) McDonald criteria for multiple sclerosis (MS). Results: Of the 21 patients, satisfying the criteria for NMO and for whom the test results were available, 17 were positive for NMO-IgG (80.9%), and of the 11 patients having LETM, 6 (54.5%) were positive for NMO-IgG. In one patient with NMO, the test result was not available. None of the 12 patients satisfying McDonald criteria for MS showed NMO-IgG seropositivity. Conclusion: Our study suggests that it is worthwhile to pursue NMO-IgG testing as a diagnostic tool for patients with clinical and Magnetic Resonance Imaging (MRI) features consistent with NMO and LETM in the Indian population

Influence of HLA-DRB1 alleles on Th1 and Th2 cytokine response to Mycobacterium tuberculosis antigens in pulmonary tuberculosis

The influence of human leukocyte antigens (HLA) on the immune response is well established. We investigated the regulatory role of HLA-DRB1 alleles on cytokine response to live M. tuberculosis and its culture filtrate antigen (CFA) in normal healthy subjects (NHS) and pulmonary tuberculosis (PTB) patients. Th1 (IFN-g and IL-12p40), Th2 (IL-4 and IL-5), pro-inflammatory (IL-6 and IL-8) and anti-inflammatory (TGF-b and IL-10) cytokines were measured by ELISA in 72-h-old peripheral blood mononuclear cell culture supernatants from 58 NHS and 48 PTB patients. HLA-DRB1 genotyping was carried out by polymerase chain reaction and dot-blot hybridization with biotinylated sequence-specific oligonucleotide probes and detection by chemiluminescence. In response to live M. tuberculosis and CFA, significantly increased levels of IL-6, IL-8 and TGF-b and decreased IFN-g, IL-12p40 and IL-10 were seen in PTB patients compared to NHS. We observed a significantly increased IFN-g response in HLA-DRB1*03-positive NHS (p ¼ 0.03) and decreased IFN-g response in HLA-DRB1*15-positive patients (p ¼ 0.04) than respective allele-negative individuals. An increased level of IL-12p40 in DRB1*10 (p ¼ 0.02) and IL-10 in DRB1*12- (p ¼ 0.03) positive NHS and an increased level of IL-6 in DRB1*04- (p ¼ 0.02) positive patients were observed. The study suggests that HLA-DRB1 alleles differentially modulate the various cytokine responses to M. tuberculosis antigens, which may influence the cellular and humoral immune responses to M. tuberculosis infection in a susceptible host

Risk factors of hypoglycaemia in elderly diabetic patients: A case–Control study from a Tertiary Hospital

Background: Hypoglycaemia is a side effect of strict diabetes control, especially in the geriatric population above sixty, who constitute approximately 100 million of the Indian population. This study was undertaken to explore the risk factors of hypoglycaemia in elderly inpatients. Materials and Methods: Fifty patients who were found to have hypoglycaemia either at admission or while admitted were enrolled. Their risk factors were compared with fifty age- and sex-matched inpatients admitted to medical wards who did not experience hypoglycaemia. Results: The duration of diabetes was significantly longer (13.4 ± 9.2 vs. 8.1 ± 5.7 years; P = 0.012) in the group which experienced hypoglycaemia. The mean glycated haemoglobin was significantly lower in the group which experienced hypoglycaemia (6.33 ± 1.12 vs. 7.61 ± 1.17; P = 0.002). Of the 50 patients who developed hypoglycaemia 28 were asymptomatic. Infection and renal failure were significantly higher in the study group. On multivariate analysis, infection was the only significant precipitating factor. Conclusion: With strict blood glucose control, elderly patients are at high risk of hypoglycaemia. The risk is higher if the patients have renal failure and infection. Diabetic therapy in elderly people should be adjusted in such a way to prevent hypoglycaemia

Cytokine gene polymorphisms and cytokine levels in pulmonary tuberculosis

Polymorphisms in the cytokine genes are known to influence cytokine levels and may be associated with outcome of infections. We investigated the polymorphisms in the cytokine genes namely IFN-c (+874 and +5644), IL-2 (�330 and +160), IL-4 (VNTR), IL-6 (�174), IL-10 (�1082 and �819) and IL-12B (+1188) in 188 normal healthy subjects (NHS) and 166 pulmonary tuberculosis patients (PTB) using polymerase chain reaction-based methods. To study the influence of cytokine gene polymorphisms on cytokine levels, phytohaemagglutinin and culture filtrate antigen of Mycobacterium tuberculosis-induced cytokine levels were measured by ELISA from 72-h-old peripheral blood mononuclear cell culture supernatants. Significantly decreased frequency of TT genotype of IL-2 �330 polymorphism (p = 0.024, odds ratio (OR) 0.53, 95% CI 0.31–0.92) was observed in patients compared to NHS. The genotype frequencies of other polymorphisms were not different between patients and NHS. IL-12p40 levels were significantly decreased among NHS with AA genotype of IL-12B gene polymorphism compared to NHS with AC genotype (p < 0.05). Increased levels of IL-12p40 were observed among patients with CC genotype of IL-12B gene compared to patients with other genotypes (p < 0.01). The present study suggests that the TT genotype of IL-2 �330 polymorphism may be associated with the protection to PTB in south India. Further, +1188 polymorphism of IL-12B gene either alone or in combination with closely linked genes may regulate IL-12p40 production and may play a major role on acquired immunity to tuberculosis